An antisense oligodeoxyribonucleotide to m-calpain mRNA inhibits secretion from alveolar epithelial type II cells.
We investigated the effect of translational suppression of m-calpain on [3H]-phosphatidylcholine (PC) secretion utilising an antisense oligodexoyribonucleotide (oligo) directed against mRNA encoding m-calpain catalytic subunit. Two types of oligo, sense (S) and antisense (AS), to a portion of exon 12 of rat m-calpain catalytic subunit mRNA were tested. Constitutive secretion was decreased by 23% by AS-oligo (1 microM) treatment, while S-oligo (1 microM) had no effect. TPA-stimulated secretion was inhibited about 50-60% by AS-oligo (1-3 microM) and the inhibition was concentration-dependent, while S-oligo (1 microM) only inhibited about 10% of TPA-stimulated secretion. Northern and Western blot analyses revealed that the AS-oligo treatment reduced m-calpain mRNA and protein levels by 32% and 78%, respectively. The data indicate that antisense strategy is effective in suppressing calpain expression and type II cell secretion.
Li
H L
HL
Institute for Environmental Medicine, University of Pennsylvania Medical Center, Philadelphia 19104, USA.
Feinstein
S I
SI
Liu
L
L
Zimmerman
U J
UJ
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002154:Calpain / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000187:drug effects; D008297:Male; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000378:metabolism; D011650:Pulmonary Alveoli / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016372:RNA, Antisense / Q000494:pharmacology; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley
D016376:Oligonucleotides, Antisense; D010713:Phosphatidylcholines; D016372:RNA, Antisense; D012333:RNA, Messenger; D002154:Calpain